The synthesis of nano silver-graphene oxide system and its efficacy against endodontic biofilms using a novel tooth model

OBJECTIVE: The deleterious caustic effects of sodium hypochlorite (NaOCl) as a root canal irrigant makes it imperative that alternative methods are developed for root canal disinfection. The purpose of this study was to examine the antimicrobial efficacy of silver nanoparticles (AgNPs) synthesized on an aqueous graphene oxide (GO) matrix (Ag-GO), with different irrigant delivery methods to enhance the disinfection regimen, using a novel ex vivo infected tooth model.

METHODS: AgNPs were prepared by reducing AgNO3 with 0.01M NaBH4 in presence of GO. Elemental analysis was performed with scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS) and scanning transmission electron microscopy (STEM) was used for size and morphology analysis of GO and Ag-GO. Nutrient stressed, multi-species biofilms were grown in prepared root canals of single-rooted teeth. The irrigants used were sterile saline, 1% and 2.5% NaOCl, 2% chlorhexidine gluconate (CHX), 17% EDTA and an aqueous suspension of 0.25% Ag-GO. The antimicrobial efficacy of the irrigants were performed with paper point sampling and measurement of microbial counts. The biofilm disruption in dentine tubule surfaces was analysed with confocal laser scanning microscopy (CLSM). The acquisition of total biovolume (μm3/μm2) and biofilm viability was performed using software BioImage_L. Two-way analysis of variance (ANOVA) with post hoc Tukey tests was used for data analysis with level of statistical significance set at P<0.05.

RESULTS: SEM/EDS analysis confirmed impregnation of Ag within the GO matrix. TEM images showed polygonal GO sheets and spherical AgNPs of diameter 20-50nm, forming a network on the surface of GO sheets. The use of ultrasonic activation enhanced the efficacy of Ag-GO compared to 1% NaOCl, 2% CHX, 17% EDTA and sterile saline (P<0.05). The microbial killing efficacy of 2.5% NaOCl was superior compared to the experimental groups. The maximum biofilm disruption, in dentine tubule surfaces, was achieved by 2.5% NaOCl, however Ag-GO caused a significant reduction of total biovolumes compared to the rest of the experimental groups (P<0.05%).

SIGNIFICANCE: The successful documentation of the microbial killing and biofilm disruption capacity of Ag-GO is a promising step forward to explore its unique properties in clinical applications and biomaterials in dentistry.